| Background Severe acute pancreatitis(SAP) is a common clinical danger disease.The mortality of severe acute pancreatitis remains increase.At the beginning acute pancreatitis is thought to be determined and mediated by a variety of pro-and anti-inflammatory mediators released from the pancreas and various other sources during the course of the disease. Currently,it is found that the degree of macrophages apoptosis is inversely related to the severity of acute pancreatitis,which suggests that the apoptosis may be a teleologically beneficial response to acute pancreatitis.So it is important to recognize the mechanism of macrophages apoptosis in severe acute pancreatitis.Objective:Severe acute pancreatitis has seriously endangered the life and health of human beings,and it is often accompanied with systemic inflammatory response syndrome(SIRS),serious infections,septic shock and multiple organ dysfunction syndrome(MODS),which lead to death of patients.With the deep study on the pathogenesis of SAP mechanism,it is demonstrated that macrophages play an important role in antigen presentation-activated immune responses of SAP patients. This paper is to explore the preparation of Clodronate-liposome and the influence of function to alveolar macrophage s(AM) in SAP rat.Methods:Clodronate-liposome is prepared by means of thin film,Eight SD rats are made into SAP model by the method of pancreatic subcapsular injection using 5%Sodium taurocholate successfully.The AMs were isolated and purified then incubated from brochoalveolar lavage by various time stick cliff test.Then it is divided into five groups including control group,SAP+ blank liposome group(50μl,100μl),SAP + Clodronate-liposome group(50μl,100μl).The AMs and different drugs are incubated at culture boards in different time.The toxiocity percent of AMs are identified by MTT.AO fluorescence and haematoxylin dye are employed to determine the apoptosis of the AMs.Result:the prepared Clodronate-liposome has a suitable encapsulation efficiency of clodronate(5.8%) with an average size of 200nm,the spherical shape of liposome is confirmed by transmission electron microscope.The AM has a normal shape,the purity is more than 90 percent determined by Wright's staining,trypan blue staining find that the rate of cell survival is more than 95 percent.With the different influence of different Clodronate-liposome doses,the MTT show that there are no significant differences between control group and blank liposome group(50μl,100μl),significant differences between control group and Clodronate-liposome group(50μl,100μl)(P＜0.01 or P＜0.05).The ODs are decreasing when the action time and doses are increasing.This show that the AMs are inhibited.Under AO fluorescence,the toxicity percent of control group was 2.08±0.73%,ANP+liposome group(3.25±0.52%,3.66±0.51%),ANP+Lipsomal clodronate group((20.17±1.72%,25.08±1.31)%,(P＜0.01).Apoptosis ratio is relation to follow the action time and Clodronate-liposome doses.AO fluorescence and haematoxylin dye are available to determine the apoptosis of the AMs.Conclusion:We found that it was feasible to prepare Clodronateliposome by means of thin film,and we prepared Clodronate-liposome successfully,it had a suitable encapsulation efficiency of clodronate with an average size confirmed by transmission electron microscope.We found that Clodronate-liposome could induce apoptosis of AMs in SAP rat.The paper provides a new theory to treat SAP in clinic.
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